Nanomaterial-based artificial enzymes (Nanozymes) have attracted recent attention because of their unique advantageous characteristics such as excellent robustness and stability, low-cost production by facile scale-up, and longterm preservation capability that are critically required as an alternative to natural enzymes. These nanozymes exhibit natural enzyme-like activity, and they have been applied to diverse kinds of detection methods for disease-associated biomolecules such as DNAs, proteins, cells, and small molecules including glucose. To highlight the progress in the field of disease diagnostics using nanozyme, this review discusses many nanozyme-based detection methods categorized by the types of target biomolecules. Finally, we address the current challenges and perspectives for the widespread utilization of nanozyme-based disease diagnostics.